Vaccination of Koalas with a Recombinant Chlamydia pecorum Major Outer Membrane Protein Induces Antibodies of Different Specificity Compared to Those Following a Natural Live Infection

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Abstract

Chlamydial infection in koalas is common across the east coast of Australia and causes significant morbidity, infertility and mortality. An effective vaccine to prevent the adverse consequences of chlamydial infections in koalas (particularly blindness and infertility in females) would provide an important management tool to prevent further population decline of this species. An important step towards developing a vaccine in koalas is to understand the host immune response to chlamydial infection. In this study, we used the Pepscan methodology to identify B cell epitopes across the Major Outer Membrane Protein (MOMP) of four C. pecorum strains/genotypes that are recognized, either following (a) natural live infection or (b) administration of a recombinant MOMP vaccine. Plasma antibodies from the koalas naturally infected with a C. pecorum G genotype strain recognised the epitopes located in the variable domain (VD) four of MOMP G and also VD4 of MOMP H. By comparison, plasma antibodies from an animal infected with a C. pecorum F genotype strain recognised epitopes in VD1, 2 and 4 of MOMP F, but not from other genotype MOMPs. When Chlamydia-free koalas were immunised with recombinant MOMP protein they produced antibodies not only against epitopes in the VDs but also in conserved domains of MOMP. Naturally infected koalas immunised with recombinant MOMP protein also produced antibodies against epitopes in the conserved domains. This work paves the way for further refinement of a MOMP-based Chlamydia vaccine that will offer wide cross-protection against the variety of chlamydial infections circulating in wild koala populations. © 2013 Kollipara et al.

Figures

  • Table 1. List of diseased as well as healthy koalas in this study with their qPCR status, infecting strain and clinical disease observed during the study.
  • Figure 1. Layout of C. pecorum MOMP peptide library. Peptides 1 to 63 represent five conserved domains (straight lines) interspersed by four variable domains (boxes) of MOMP F. Peptides 64 to 88 represent four variable domains of MOMP A, MOMP G and MOMP H, respectively.
  • Figure 2. B cell epitope mapping across four MOMPs in Chlamydia free koalas after vaccination. Response profiles and immunoblots of four Chlamydia free koalas vaccinated with MOMP A (A), MOMP F (B), MOMP G (C) and MOMP A as well as MOMP F (D), respectively. The blue line indicates the absorbance of pre immunized samples collected from individual Chlamydianegative animals. The red line indicates the 20 weeks post immunized sample. The green line indicates the background (Mean and two times standard deviation of negative controls).
  • Figure 3. B cell epitope mapping across four MOMPs in naturally, C. pecorum genotype G infected koalas. Response profiles of two wild koalas naturally infected with C. pecorum G, Popeye (A) and Mars Bar (B) across MOMP peptide library and amino acid alignment of four MOMPs at VD4 epitope (C) (dots represent the consensus). Representative blot of plasma from the koalas infected with C. pecorum G against four recombinant full length MOMP proteins (D). The blue line indicates the mean of absorbance of four Chlamydia free animals. The red line indicates the Chlamydia infected animal. The green line indicates the background (Mean and two times standard deviation of negative controls).
  • Figure 4. B cell epitope mapping across four MOMPs in naturally, C. pecorum genotype F infected koalas. Response profiles of two wild koalas naturally infected with C. pecorum F, Nixon (A) and Felix Pitt (B) across MOMP peptide library. Alignment of four MOMPs at an epitope at variable domain two (C), and two overlapping epitopes at variable domain four (D) (dots represent the consensus). The blue line indicates the mean of absorbance of four Chlamydia free animals. The red line indicates the Chlamydia infected animal. The green line indicates the background (Mean and two times standard deviation of negative controls).
  • Figure 5. B cell epitope mapping across four MOMPs in Chlamydia infected koalas after administration with placebo or vaccine. Response profile of a C. pecorum F infected koala administered with a placebo (adjuvant only) across our MOMP peptide library (A). Response profiles of a naturally infected C. pecorum G koala immunized with recombinant MOMP G (B), and a naturally infected C. pecorum F koala immunized with recombinant MOMP G (C) across our MOMP peptide library. The blue line represents the absorbance of the pre-immunized sample. The red line represents the 20 weeks post-immunized sample. The green line indicates the background (Mean and two times standard deviation of negative controls).

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APA

Kollipara, A., Polkinghorne, A., Beagley, K. W., & Timms, P. (2013). Vaccination of Koalas with a Recombinant Chlamydia pecorum Major Outer Membrane Protein Induces Antibodies of Different Specificity Compared to Those Following a Natural Live Infection. PLoS ONE, 8(9). https://doi.org/10.1371/journal.pone.0074808

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